Structural, thermodynamic and kinetic studies of antibo*

抗体的结构、热力学和动力学研究*

• 批准号: 6644845
• 负责人: BRADFORD C BRADEN
• 金额: $ 3.74万
• 依托单位: BOWIE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6644845
• 关键词: X ray crystallography; antiantibody; antigen antibody reaction; antiidiotype antibody; binding sites; cell component structure /function; chemical kinetics; cooperative study; fluorescence polarization; hybridomas; immunoglobulin G; immunoglobulin M; immunoglobulins; intermolecular interaction; laboratory mouse; lysozyme; microcalorimetry; molecular cloning; monoclonal antibody; nucleic acid sequence; protein binding; stop flow technique; structural biology; surface plasmon resonance; thermodynamics; ultracentrifugation

DESCRIPTION (provided by applicant) Affinity maturation of the immune response is a process unique to the immune system. This process consists of somatic hypermutation of the variable domains of immunoglobulins, which leads to a significant increase in the affinities of the antibodies elicited during a humoral immune response. The understanding of the molecular and structural basis of this phenomenon is an area of particular interest for Immunology as well as the biological sciences in general. Several authors have described, in detail, models of affinity maturation in antibody-hapten systems. However, affinity maturation against protein antigens has not been described in such detail. Our previous work, as in other laboratories, has shown that the antibodies elicited during the immune response against a protein antigen show a fast maturation of the affinity, a phenomenon closely related to the early IgM-IgG isotype class switch process. No structural, sequence or functional studies describing the changes that occur at the combining site of anti-protein antibodies during this process can be found in the current literature. The aim of this project is to characterize the sequence and the binding site structures of the IgM unmutated precursors of the structurally well characterized IgG anti-hen egg-white lysozyme (HEL) antibodies. This goal will be accomplished by cloning and sequencing anti-HEL IgMs and by X-ray crystallographic structure determinations of the Fv fragments of these IgMs. Structural, functional and mutational studies of the recombinant Fv fragments of those IgMs would reveal the nature of the changes produced during affinity maturation.

描述（由申请人提供） 免疫反应的亲和力成熟是免疫独特的过程 系统。该过程由可变域的体细胞超突变组成 免疫球蛋白，这导致亲和力显着增加 体液免疫反应过程中引发的抗体。的理解 这种现象的分子和结构基础是一个特殊的领域 对免疫学以及一般生物科学的兴趣。 几位作者详细描述了亲和力成熟模型 抗体-半抗原系统。然而，针对蛋白质抗原的亲和力成熟 还没有如此详细地描述。我们之前的工作，和其他工作一样 实验室表明，在免疫反应过程中引发的抗体 针对蛋白质抗原表现出亲和力快速成熟的现象 与早期 IgM-IgG 同种型转换过程密切相关。不 结构、序列或功能研究描述了发生的变化 在此过程中可以找到抗蛋白抗体的结合位点 在当前的文献中。 该项目的目的是表征序列和结合位点 结构良好的 IgM 未突变前体的结构 表征了 IgG 抗鸡蛋清溶菌酶 (HEL) 抗体。这个目标将 通过抗 HEL IgM 的克隆和测序以及 X 射线来完成 这些 IgM 的 Fv 片段的晶体结构测定。 重组Fv片段的结构、功能和突变研究 这些 IgM 的数量将揭示亲和过程中产生的变化的本质 成熟。